論文 - 詳細
| RRC ID | 10877 |
|---|---|
| 著者 | Shah A, Farooq AV, Tiwari V, Kim MJ, Shukla D. |
| タイトル | HSV-1 infection of human corneal epithelial cells: receptor-mediated entry and trends of re-infection. |
| ジャーナル | Mol Vis |
| Abstract |
PURPOSE:The human cornea is a primary target for herpes simplex virus-1 (HSV-1) infection. The goals of the study were to determine the cellular modalities of HSV-1 entry into human corneal epithelial (HCE) cells. Specific features of the study included identifying major entry receptors, assessing pH dependency, and determining trends of re-infection. METHODS:A recombinant HSV-1 virus expressing beta-galactosidase was used to ascertain HSV-1 entry into HCE cells. Viral replication within cells was confirmed using a time point plaque assay. Lysosomotropic agents were used to test for pH dependency of entry. Flow cytometry and immunocytochemistry were used to determine expression of three cellular receptors--nectin-1, herpesvirus entry mediator (HVEM), and paired immunoglobulin-like 2 receptor alpha (PILR-a). The necessity of these receptors for viral entry was tested using antibody-blocking. Finally, trends of re-infection were investigated using viral entry assay and flow cytometry post-primary infection. RESULTS:Cultured HCE cells showed high susceptibility to HSV-1 entry and replication. Entry was demonstrated to be pH dependent as blocking vesicular acidification decreased entry. Entry receptors expressed on the cell membrane include nectin-1, HVEM, and PILR-α. Receptor-specific antibodies blocked entry receptors, reduced viral entry and indicated nectin-1 as the primary receptor used for entry. Cells re-infected with HSV-1 showed a decrease in entry, which was correlated to decreased levels of nectin-1 as demonstrated by flow cytometry. CONCLUSIONS:HSV-1 is capable of developing an infection in HCE cells using a pH dependent entry process that involves primarily nectin-1 but also the HVEM and PILR-α receptors. Re-infected cells show decreased levels of entry, correlated with a decreased level of nectin-1 receptor expression. |
| 巻・号 | 16 |
| ページ | 2476-86 |
| 公開日 | 2010-11-20 |
| PII | 265 |
| PMID | 21139972 |
| PMC | PMC2994737 |
| MeSH | Antibodies, Blocking / immunology Cell Adhesion Molecules / genetics Cell Adhesion Molecules / metabolism Cells, Cultured Down-Regulation / genetics Epithelial Cells / virology* Epithelium, Corneal / cytology* Flow Cytometry Fluorescent Antibody Technique Herpesvirus 1, Human / physiology* Humans Hydrogen-Ion Concentration Keratitis, Herpetic / pathology Keratitis, Herpetic / virology* Nectins Receptors, Virus / metabolism* Reverse Transcriptase Polymerase Chain Reaction Virus Internalization* Virus Replication / physiology |
| IF | 2.202 |
| 引用数 | 37 |
| WOS 分野 | OPHTHALMOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| ヒト・動物細胞 | HCE-T(RCB2280) |